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ENVIRONMENTAL STRESS AND ADAPTATION TO STRESS

Nitrite Reduces Cytoplasmic Acidosis under Anoxia¹

Department of Plant Sciences, University of Oxford, Oxford OX1 3RB, United Kingdom (I.G.L.L., M.D.F., R.G.R.); and Department of Systems Ecology, Vrije Universiteit Amsterdam, NL–1081 HV Amsterdam, The Netherlands (P.M.v.B.)

The ameliorating effect of nitrate on the acidification of the cytoplasm during short-term anoxia was investigated in maize (Zea mays) root segments. Seedlings were grown in the presence or absence of nitrate, and changes in the cytoplasmic and vacuolar pH in response to the imposition of anoxia were measured by in vivo ³¹P nuclear magnetic resonance spectroscopy. Soluble ions and metabolites released to the suspending medium by the anoxic root segments were measured by high-performance liquid chromatography and ¹H nuclear magnetic resonance spectroscopy, and volatile metabolites were measured by gas chromatography and gas chromatography-mass spectrometry. The beneficial effect of nitrate on cytoplasmic pH regulation under anoxia occurred despite limited metabolism of nitrate under anoxia, and modest effects on the ions and metabolites, including fermentation end products, released from the anoxic root segments. Interestingly, exposing roots grown and treated in the absence of nitrate to micromolar levels of nitrite during anoxia had a beneficial effect on the cytoplasmic pH that was comparable to the effect observed for roots grown and treated in the presence of nitrate. It is argued that nitrate itself is not directly responsible for improved pH regulation under anoxia, contrary to the usual assumption, and that nitrite rather than nitrate should be the focus for further work on the beneficial effect of nitrate on flooding tolerance.

² Present address: Department of Plant Biology, Michigan State University, Wilson Drive, East Lansing, MI 48824–1312.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: R.G. Ratcliffe (george.ratcliffe{at}plants.ox.ac.uk).

www.plantphysiol.org/cgi/doi/10.1104/pp.106.088898

^* Corresponding author; e-mail george.ratcliffe{at}plants.ox.ac.uk; fax 44–(0)1865–275074.

Received August 30, 2006; accepted October 19, 2006; published October 27, 2006.

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